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Methods Mol Biol DOI:10.1007/978-1-4939-8967-6_10

Genome-Wide CRISPR/Cas9 Screening for Identification of Cancer Genes in Cell Lines.

Publication TypeJournal Article
Year of Publication2019
AuthorsAdelmann, CH, Wang, T, Sabatini, DM, Lander, ES
JournalMethods Mol Biol
Volume1907
Pages125-136
Date Published2019
ISSN1940-6029
Abstract

In this protocol, pooled sgRNA libraries targeting thousands of genes are computationally designed, generated using microarray-based synthesis techniques, and packaged into lentiviral particles. Target cells of interest are transduced with the lentiviral sgRNA pools to generate a collection of knockout mutants-via Cas9-mediated genomic cleavage-and screened for a phenotype of interest. The relative abundance of each mutant in the population can be monitored over time through high-throughput sequencing of the integrated sgRNA expression cassettes. Using this technique, we outline strategies for the identification of cancer driver genes and genes mediating drug response.

DOI10.1007/978-1-4939-8967-6_10
Pubmed

http://www.ncbi.nlm.nih.gov/pubmed/30542996?dopt=Abstract

Alternate JournalMethods Mol. Biol.
PubMed ID30542996
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